CN106244475A - A kind of low temperature resistant compost fermentation microbial inoculum with deodorization functions and application thereof - Google Patents

A kind of low temperature resistant compost fermentation microbial inoculum with deodorization functions and application thereof Download PDF

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Publication number
CN106244475A
CN106244475A CN201610694440.0A CN201610694440A CN106244475A CN 106244475 A CN106244475 A CN 106244475A CN 201610694440 A CN201610694440 A CN 201610694440A CN 106244475 A CN106244475 A CN 106244475A
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microbial inoculum
bacillus
specially
aspergillus niger
aspergillus oryzae
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武凤霞
刘善江
刘建斌
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Beijing Academy of Agriculture and Forestry Sciences
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Beijing Academy of Agriculture and Forestry Sciences
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • C12N1/16—Yeasts; Culture media therefor
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D53/00—Separation of gases or vapours; Recovering vapours of volatile solvents from gases; Chemical or biological purification of waste gases, e.g. engine exhaust gases, smoke, fumes, flue gases, aerosols
    • B01D53/34—Chemical or biological purification of waste gases
    • B01D53/46—Removing components of defined structure
    • B01D53/48—Sulfur compounds
    • B01D53/50—Sulfur oxides
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D53/00—Separation of gases or vapours; Recovering vapours of volatile solvents from gases; Chemical or biological purification of waste gases, e.g. engine exhaust gases, smoke, fumes, flue gases, aerosols
    • B01D53/34—Chemical or biological purification of waste gases
    • B01D53/46—Removing components of defined structure
    • B01D53/48—Sulfur compounds
    • B01D53/52—Hydrogen sulfide
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D53/00—Separation of gases or vapours; Recovering vapours of volatile solvents from gases; Chemical or biological purification of waste gases, e.g. engine exhaust gases, smoke, fumes, flue gases, aerosols
    • B01D53/34—Chemical or biological purification of waste gases
    • B01D53/46—Removing components of defined structure
    • B01D53/54—Nitrogen compounds
    • B01D53/56—Nitrogen oxides
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D53/00—Separation of gases or vapours; Recovering vapours of volatile solvents from gases; Chemical or biological purification of waste gases, e.g. engine exhaust gases, smoke, fumes, flue gases, aerosols
    • B01D53/34—Chemical or biological purification of waste gases
    • B01D53/46—Removing components of defined structure
    • B01D53/54—Nitrogen compounds
    • B01D53/58—Ammonia
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01D—SEPARATION
    • B01D53/00—Separation of gases or vapours; Recovering vapours of volatile solvents from gases; Chemical or biological purification of waste gases, e.g. engine exhaust gases, smoke, fumes, flue gases, aerosols
    • B01D53/34—Chemical or biological purification of waste gases
    • B01D53/74—General processes for purification of waste gases; Apparatus or devices specially adapted therefor
    • B01D53/84—Biological processes
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05D—INORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C; FERTILISERS PRODUCING CARBON DIOXIDE
    • C05D9/00—Other inorganic fertilisers
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F11/00—Other organic fertilisers
    • C05F11/08—Organic fertilisers containing added bacterial cultures, mycelia or the like
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
    • C05F17/00—Preparation of fertilisers characterised by biological or biochemical treatment steps, e.g. composting or fermentation
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
    • C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
    • Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
    • Y02A50/20—Air quality improvement or preservation, e.g. vehicle emission control or emission reduction by using catalytic converters
    • Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
    • Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
    • Y02W—CLIMATE CHANGE MITIGATION TECHNOLOGIES RELATED TO WASTEWATER TREATMENT OR WASTE MANAGEMENT
    • Y02W30/00—Technologies for solid waste management
    • Y02W30/40—Bio-organic fraction processing; Production of fertilisers from the organic fraction of waste or refuse

Abstract

The invention discloses a kind of low temperature resistant compost fermentation microbial inoculum with deodorization functions and application thereof.The microorganism species contained by low temperature resistant compost fermentation microbial inoculum that the present invention provides can tolerate low temperature, can be with normal growth at 15 20 DEG C, beneficially organic materials ferments temperature in low temperature environment, mutually promotes between each strain, produce mutual synergy, thus accelerate to become thoroughly decomposed.It is experimentally confirmed: in composting process, the low temperature resistant compost fermentation microbial inoculum of the present invention is added in organic materials and ferments, not only increase the sum of compost initial stage effective microbe, accelerate compost material programming rate, shorten the anaerobic fermentation time, accelerate maturity, and reduce the discharge of Nitrogen or sulfur-compounds, optimize the physicochemical property of tunning, there is certain deodorization functions.

Description

A kind of low temperature resistant compost fermentation microbial inoculum with deodorization functions and application thereof
Technical field
The invention belongs to complex micro organism fungicide field, be specifically related to a kind of low temperature resistant compost fermentation with deodorization functions Microbial inoculum and application thereof.
Background technology
The generation amount of China's agricultural organic waste is huge, it is estimated that annual generation feces of livestock and poultry 3,000,000,000 tons, crops straw Stalk more than 700,000,000 ton, these organic wastes are rich in the nutrient needed for a large amount of crop production.Permissible by the method for aerobic compost Being high-quality fertilizer by these reclaiming organic wastes, Traditional compost method is few due to initial stage effective microbe quantity, needs Certain time could breed, and there is fermentation time long, and foul smell discharge is big, and nutrient element loss is many, serious waste of resources etc. Problem, the most northerly autumn and winter, owing to ambient temperature is low, compost fermentation plays the particularly problematic of temperature difficulty.
It addition, composting process is often along with the generation of foul gas, make people's inappetence, feel dizzy, Nausea and vomiting, Become the important restriction factor affecting compost industry development.Foul gas main component includes sulfide, Ammonia, lower aliphatic Amine, aromatic hydrocarbons, hydroxy compounds, alcohols, lower fatty acid and indole etc., wherein ammonia and hydrogen sulfide are main foul gas groups Becoming composition, the volatilization of these gases not only pollutes environment but also have lost nitrogen element, reduces the fertilizer efficiency of composting production.
Summary of the invention
First purpose of the present invention is to provide a kind of complex micro organism fungicide.
The active component of the complex micro organism fungicide that the present invention provides is Saccharomyces Cerevisiae in S accharomyces Cerevisiae, Candida maltosa Candida maltosa, bacillus subtilis Bacillus subtilis, solution starch Bacillus amyloliquefaciens, aspergillus oryzae Aspergillus oryzae, aspergillus niger Aspergillus Niger, bacstearothermophilus Bacillus stearothermophilus and sporotrichum thermophile Sporotrichum thermophile。
In above-mentioned microbial inoculum, described Saccharomyces Cerevisiae in S accharomyces cerevisiae, described Candida maltosa Candida maltosa, described bacillus subtilis Bacillus subtilis, described bacillus amyloliquefaciens Bacillus Amyloliquefaciens, described aspergillus oryzae Aspergillus oryzae, described aspergillus niger Aspergillus niger, institute State bacstearothermophilus Bacillus stearothermophilus and described sporotrichum thermophile Sporotrichum The proportioning of the cfu of thermophile is (30-60): (30-60): 500:(200-500): (20-50): (20-50): (100- 200): (20-30).
In above-mentioned microbial inoculum, described Saccharomyces Cerevisiae in S accharomyces cerevisiae, described Candida maltosa Candida maltosa, described bacillus subtilis Bacillus subtilis, described bacillus amyloliquefaciens Bacillus Amyloliquefaciens, described aspergillus oryzae Aspergillus oryzae, described aspergillus niger Aspergillus niger, institute State bacstearothermophilus Bacillus stearothermophilus and described sporotrichum thermophile Sporotrichum The proportioning of the cfu of thermophile is specially 5:5:30:50:2:2:20:2 or 5:6:50:50:5:3:10:2.
In above-mentioned microbial inoculum, described Saccharomyces Cerevisiae in S accharomyces cerevisiae is saccharomyces cerevisiae Saccharomyces cerevisiae W052502CGMCC No.12701；
Described Candida maltosa Candida maltosa is Candida maltosa Candida maltosa W052501CGMCC No.12702；
Described bacillus subtilis Bacillus subtilis is bacillus subtilis Bacillus subtilis W11025CGMCC No.12705；
Described bacillus amyloliquefaciens Bacillus amyloliquefaciens is bacillus amyloliquefaciens Bacillus amyloliquefaciens W160617CGMCC No.12703；
Described aspergillus oryzae Aspergillus oryzae is aspergillus oryzae Aspergillus oryzae W060406CGMCC No.12627；
Described aspergillus niger Aspergillus niger is aspergillus niger Aspergillus niger W30117CGMCC No.12625；
Described bacstearothermophilus Bacillus stearothermophilus is bacstearothermophilus Bacillus stearothermophilus W10208CGMCC No.12704；
Described sporotrichum thermophile Sporotrichum thermophile is sporotrichum thermophile Sporotrichum thermophile W2441CGMCC No.12626。
Second object of the present invention is to provide the preparation method of a kind of complex micro organism fungicide.
The preparation method of the complex micro organism fungicide that the present invention provides is by Saccharomyces Cerevisiae in S accharomyces Cerevisiae microbial inoculum, Candida maltosa Candida maltosa microbial inoculum, bacillus subtilis Bacillus Subtilis microbial inoculum, bacillus amyloliquefaciens Bacillus amyloliquefaciens microbial inoculum, aspergillus oryzae Aspergillus Oryzae microbial inoculum, aspergillus niger Aspergillus niger microbial inoculum, bacstearothermophilus Bacillus Stearothermophilus microbial inoculum and sporotrichum thermophile Sporotrichum thermophile microbial inoculum are uniformly mixed so as to obtain.
In said method, described Saccharomyces Cerevisiae in S accharomyces cerevisiae microbial inoculum, described maltose vacation silk ferment Female Candida maltosa microbial inoculum, described bacillus subtilis Bacillus subtilis microbial inoculum, described solution starch spore bar Bacterium Bacillus amyloliquefaciens microbial inoculum, described aspergillus oryzae Aspergillus oryzae microbial inoculum, described aspergillus niger Aspergillus niger microbial inoculum, described bacstearothermophilus Bacillus stearothermophilus microbial inoculum and institute The mass ratio stating sporotrichum thermophile Sporotrichum thermophile microbial inoculum is (1-3): (1-3): 2:(1-3): (1-3): (1-3): (1-2): (1-2).
In said method, described Saccharomyces Cerevisiae in S accharomyces cerevisiae microbial inoculum, described maltose vacation silk ferment Female Candida maltosa microbial inoculum, described bacillus subtilis Bacillus subtilis microbial inoculum, described solution starch spore bar Bacterium Bacillus amyloliquefaciens microbial inoculum, described aspergillus oryzae Aspergillus oryzae microbial inoculum, described aspergillus niger Aspergillus niger microbial inoculum, described bacstearothermophilus Bacillus stearothermophilus microbial inoculum and institute The mass ratio stating sporotrichum thermophile Sporotrichum thermophile microbial inoculum is specially 12:10:14:12:10:14:15:13 Or 12:13:8:14:15:13:8:8.
In said method, described Saccharomyces Cerevisiae in S accharomyces cerevisiae is saccharomyces cerevisiae Saccharomyces cerevisiae W052502CGMCC No.12701；
Described Candida maltosa Candida maltosa is Candida maltosa Candida maltosa W052501CGMCC No.12702；
Described bacillus subtilis Bacillus subtilis is bacillus subtilis Bacillus subtilis W11025CGMCC No.12705；
Described bacillus amyloliquefaciens Bacillus amyloliquefaciens is bacillus amyloliquefaciens Bacillus amyloliquefaciens W160617CGMCC No.12703；
Described aspergillus oryzae Aspergillus oryzae is aspergillus oryzae Aspergillus oryzae W060406CGMCC No.12627；
Described aspergillus niger Aspergillus niger is aspergillus niger Aspergillus niger W30117CGMCC No.12625；
Described bacstearothermophilus Bacillus stearothermophilus is bacstearothermophilus Bacillus stearothermophilus W10208CGMCC No.12704；
Described sporotrichum thermophile Sporotrichum thermophile is sporotrichum thermophile Sporotrichum thermophile W2441CGMCC No.12626。
In said method,
The preparation method of described Saccharomyces Cerevisiae in S accharomyces cerevisiae microbial inoculum is as follows: by saccharomyces cerevisiae Saccharomyces cerevisiae W052502 inoculation is in YPD culture medium, at 29 DEG C, trains under conditions of 200rmp Support 20 hours, then after 3000r/min centrifugal concentrating 15-20min, with zeolite powder according to the ratio (tool that mass ratio is 1:9-1:5 Body is 1:7) absorption mixing, obtain Saccharomyces Cerevisiae in S accharomyces cerevisiae microbial inoculum.Wherein, saccharomyces cerevisiae The concentration of Saccharomyces cerevisiae W052502 bacterial strain is 5,000,000,000 cfu/g.
The preparation method of described Candida maltosa Candida maltosa microbial inoculum is: by Candida maltosa Candida maltosa W052501 inoculation is in YPD culture medium, at 30 DEG C, and cultivation 22 hours under the conditions of 200rmp, so After rear 3000r/min centrifugal concentrating 15-20min, inhale according to the ratio (specially 1:7) that mass ratio is 1:9-1:5 with zeolite powder Attached mixing, obtains Candida maltosa Candida maltosa microbial inoculum.Wherein, Candida maltosa Candida The concentration of maltosa W052501 bacterial strain is 6,000,000,000 cfu/g.
The preparation method of described bacillus subtilis Bacillus subtilis microbial inoculum is: by bacillus subtilis In Bacillus subtilis W11025 inoculation beef-protein medium, at 30 DEG C, cultivate under the conditions of 230rmp Reach 85% to spore forming rate, then after 3000r/min centrifugal concentrating 15-20min, be 1:9-1 with zeolite powder according to mass ratio: Ratio (specially 1:5) the absorption mixing of 2, obtains bacillus subtilis Bacillus subtilis microbial inoculum.Wherein, hay bud The concentration of spore bacillus Bacillus subtilis W11025 bacterial strain is 50,000,000,000 cfu/g.
The preparation method of described bacillus amyloliquefaciens Bacillus amyloliquefaciens microbial inoculum is: will solve starch Bacillus amyloliquefaciens W160617 inoculation is in beef-protein medium, 31 DEG C, cultivate under the conditions of 230rmp to spore forming rate and reach 85%, then after 3000r/min centrifugal concentrating 15-20min, with zeolite Powder, according to ratio (specially 1:5) the absorption mixing that mass ratio is 1:9-1:2, obtains bacillus amyloliquefaciens Bacillus Amyloliquefaciens microbial inoculum.Wherein, bacillus amyloliquefaciens Bacillus amyloliquefaciens W160617 bacterium The concentration of strain is 50,000,000,000 cfu/g.
The preparation method of described aspergillus oryzae Aspergillus oryzae microbial inoculum is: by aspergillus oryzae Aspergillus Oryzae W060406 inoculation is in PDA culture medium, at 28 DEG C, cultivates 18h, then 3000r/min under the conditions of 100rmp After centrifugal concentrating 15-20min, with zeolite powder according to ratio (specially 1:5) the absorption mixing that mass ratio is 1:9-1:2, obtain Aspergillus oryzae Aspergillus oryzae microbial inoculum.Wherein, the concentration of aspergillus oryzae Aspergillus oryzae W060406 bacterial strain It is 5,000,000,000 cfu/g.
The preparation method of described aspergillus niger Aspergillus niger microbial inoculum is: by aspergillus niger Aspergillus niger W30117 inoculation is in PDA culture medium, at 32 DEG C, cultivates 20h, then 3000r/min centrifugal concentrating under the conditions of 100rmp After 15-20min, with zeolite powder according to ratio (specially 1:5) the absorption mixing that mass ratio is 1:9-1:2, obtain aspergillus niger Aspergillus niger microbial inoculum.Wherein, the concentration of aspergillus niger Aspergillus niger W30117 bacterial strain is 3,000,000,000 cfu/ g。
The preparation method of described bacstearothermophilus Bacillus stearothermophilus microbial inoculum is: will be addicted to Hot Bacillus stearothermophilus Bacillus stearothermophilus W10208 inoculation is in beef-protein medium In, at 40 DEG C, cultivate under the conditions of 230rmp to spore forming rate and reach 85%, then after 3000r/min centrifugal concentrating 15-20min, With zeolite powder according to ratio (specially 1:5) the absorption mixing that mass ratio is 1:9-1:2, obtain bacstearothermophilus Bacillus stearothermophilus microbial inoculum.Wherein, bacstearothermophilus Bacillus The concentration of stearothermophilus W10208 bacterial strain is 10,000,000,000 cfu/g.
The preparation method of described sporotrichum thermophile Sporotrichum thermophile microbial inoculum is: by sporotrichum thermophile Sporotrichum thermophile W2441 inoculation is in PDA culture medium, at 45 DEG C, cultivates under the conditions of 100rmp 20h, then after 3000r/min centrifugal concentrating 15-20min, with zeolite powder according to the ratio that mass ratio is 1:9-1:2 (specially 1:5) absorption mixing, obtains sporotrichum thermophile Sporotrichum thermophile microbial inoculum.Wherein, sporotrichum thermophile The concentration of Sporotrichum thermophile W2441 bacterial strain is 2,000,000,000 cfu/g.
Third object of the present invention is to provide the new application of above-mentioned complex micro organism fungicide.
The invention provides the application in following (1)-(8) in any one of the above-mentioned complex micro organism fungicide:
(1) compost fermentation agent or decomposing agent are prepared；
(2) compost under compost, particularly cryogenic conditions；
(3) organic materials or the fermentation of organic waste；
(4) programming rate of fermentation heap body is accelerated；
(5) discharge of odorous gas in digest process is reduced；
(6) plant height of plant and/or root length and/or root fresh weight and/or root dry weight and/or yield are improved；
(7) the malformed fruit rate of Tree Fruit is reduced；
(8) single fruit weight of Tree Fruit and/or sugar content and/or yield are improved；
Described odorous gas is hydrogen sulfide and/or sulfur dioxide and/or nitrogen dioxide and/or ammonia.
Fourth object of the present invention is to provide a kind of decomposing agent for compost.
The decomposing agent that the present invention provides is made up of with carrier above-mentioned complex micro organism fungicide.
In above-mentioned decomposing agent, described carrier is the mixture of powdered rice hulls or zeolite powder or powdered rice hulls and zeolite powder.
In above-mentioned decomposing agent, the fineness of described carrier is more than or equal to 60 mesh.
In above-mentioned decomposing agent, described complex micro organism fungicide is (49-201) with the mass ratio of described carrier: 909.
In above-mentioned decomposing agent, the mass ratio of described complex micro organism fungicide and described carrier is specially 49:451 or 91: 909。
5th purpose of the present invention is to provide the new application of above-mentioned decomposing agent.
The invention provides above-mentioned decomposing agent following 1)-7) in application in any one:
1) compost fermentation under compost, particularly cryogenic conditions；
2) organic materials or the fermentation of organic waste；
3) programming rate of fermentation heap body is accelerated；
4) discharge of odorous gas in digest process is reduced；
5) plant height of plant and/or root length and/or root fresh weight and/or root dry weight and/or yield are improved；
6) the malformed fruit rate of Tree Fruit is reduced；
7) single fruit weight of Tree Fruit and/or sugar content and/or yield are improved；
Described odorous gas is hydrogen sulfide and/or sulfur dioxide and/or nitrogen dioxide and/or ammonia.
6th a kind of method that purpose is to provide compost of the present invention.
The method of the compost that the present invention provides comprises the steps: to process organic materials or organic waste with above-mentioned decomposing agent Thing.
In said method, described decomposing agent is (1-5) with the mass ratio of described organic materials or organic waste: 1000.
In said method, described organic materials can be, but not limited to fowl and animal excrement, agricultural crop straw, vegetable tail dish straw, Mushroom residue, furfural dregs, mud etc..
7th purpose of the present invention is to provide the preparation method of a kind of organic fertilizer.
The preparation method of organic fertilizer that the present invention provides comprise the steps: with above-mentioned decomposing agent process organic materials or Organic waste, obtains organic fertilizer.
In said method, described decomposing agent is (1-5) with the mass ratio of described organic materials or organic waste: 1000.
In said method, described organic materials can be, but not limited to fowl and animal excrement, agricultural crop straw, vegetable tail dish straw, Mushroom residue, furfural dregs, mud etc..
The organic fertilizer that said method prepares falls within protection scope of the present invention.
Final object of the present invention is to provide the new application of above-mentioned organic fertilizer.
The invention provides above-mentioned organic fertilizer at following a1)-a3) in application in any one:
A1) plant height of plant and/or root length and/or root fresh weight and/or root dry weight and/or yield are improved；
A2) the malformed fruit rate of Tree Fruit is reduced；
A3) single fruit weight of Tree Fruit and/or sugar content and/or yield are improved.
The invention provides a kind of compost composite ferment, its contained microorganism species can tolerate low temperature, at 15-20 DEG C can be with normal growth, beneficially organic materials ferments temperature in low temperature environment, mutually promotes between each strain, produces mutually association Same effect, thus accelerate to become thoroughly decomposed.It is experimentally confirmed: in composting process, the compost composite ferment of the present invention is added to Organic materials ferments, not only increases the sum of compost initial stage effective microbe, accelerate compost material programming rate, Shorten the anaerobic fermentation time, accelerate maturity, and reduce the discharge of Nitrogen or sulfur-compounds, optimize tunning Physicochemical property, there is certain deodorization functions.
Accompanying drawing explanation
Fig. 1 is the variations in temperature of the fermentation heap body in embodiment 2.Wherein, abscissa is the time (h), and vertical coordinate is temperature (℃)。
Fig. 2 is the variations in temperature of the fermentation heap body in embodiment 3.Wherein, abscissa is the time (h), and vertical coordinate is temperature (℃)。
Fig. 3 is form and the Physiology and biochemistry qualification result of bacterial strain W11025.
Fig. 4 is the sequence of the 16s rDNA of bacterial strain W11025.
Fig. 5 is cellular morphology and the Physiology and biochemistry qualification result of bacterial strain W160617.
Fig. 6 is the sequence of the 16s rDNA of bacterial strain W11025.
Fig. 7 is the physicochemical characteristics of bacterial strain W052501.
Fig. 8 is the rRNA gene D1D2 region sequence of bacterial strain W052501.
Fig. 9 is the sequence of the ITS sequence of bacterial strain W30117.
Preservation explanation
Strain name: saccharomyces cerevisiae
Latin name: Saccharomyces cerevisiae
Strain number: W052502
Preservation mechanism: China Committee for Culture Collection of Microorganisms's common micro-organisms center
Preservation mechanism is called for short: CGMCC
Address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Preservation date: on June 22nd, 2016
Register on the books numbering in preservation center: CGMCC No.12701
Strain name: Candida maltosa
Latin name: Candida maltosa
Strain number: W052501
Preservation mechanism: China Committee for Culture Collection of Microorganisms's common micro-organisms center
Preservation mechanism is called for short: CGMCC
Address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Preservation date: on June 22nd, 2016
Register on the books numbering in preservation center: CGMCC No.12702
Strain name: bacillus subtilis
Latin name: Bacillus subtilis
Strain number: W11025
Preservation mechanism: China Committee for Culture Collection of Microorganisms's common micro-organisms center
Preservation mechanism is called for short: CGMCC
Address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Preservation date: on June 22nd, 2016
Register on the books numbering in preservation center: CGMCC No.12705
Strain name: bacillus amyloliquefaciens
Latin name: Bacillus amyloliquefaciens
Strain number: W160617
Preservation mechanism: China Committee for Culture Collection of Microorganisms's common micro-organisms center preservation mechanism is called for short: CGMCC
Address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Preservation date: on June 22nd, 2016
Register on the books numbering in preservation center: CGMCC No.12703
Strain name: aspergillus oryzae
Latin name: Aspergillus oryzae
Strain number: W060406
Preservation mechanism: China Committee for Culture Collection of Microorganisms's common micro-organisms center preservation mechanism is called for short: CGMCC
Address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Preservation date: on June 22nd, 2016
Register on the books numbering in preservation center: CGMCC No.12627
Strain name: aspergillus niger
Latin name: Aspergillus niger
Strain number: W30117
Preservation mechanism: China Committee for Culture Collection of Microorganisms's common micro-organisms center preservation mechanism is called for short: CGMCC
Address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Preservation date: on June 22nd, 2016
Register on the books numbering in preservation center: CGMCC No.12625
Strain name: bacstearothermophilus
Latin name: Bacillus stearothermophilus
Strain number: W10208
Preservation mechanism: China Committee for Culture Collection of Microorganisms's common micro-organisms center preservation mechanism is called for short: CGMCC
Address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Preservation date: on June 22nd, 2016
Register on the books numbering in preservation center: CGMCC No.12704
Strain name: sporotrichum thermophile
Latin name: Sporotrichum thermophile
Strain number: W2441
Preservation mechanism: China Committee for Culture Collection of Microorganisms's common micro-organisms center
Preservation mechanism is called for short: CGMCC
Address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3
Preservation date: on June 22nd, 2016
Register on the books numbering in preservation center: CGMCC No.12626
Detailed description of the invention
Experimental technique used in following embodiment if no special instructions, is conventional method.
Material used in following embodiment, reagent etc., if no special instructions, the most commercially obtain.
Quantitative test in following embodiment, is respectively provided with three times and repeats experiment, results averaged.
The solvent of the YPD culture medium in following embodiment is water, and solute and mass fraction in the medium thereof are 1% ferment Female powder, 2% peptone, 2% glucose.
The solvent of the beef-protein medium in following embodiment is water, and solute and quality in the medium thereof are divided Number is 0.3% Carnis Bovis seu Bubali cream, 1% peptone, 2% glucose, 0.5% sodium chloride.
The preparation method of the PDA culture medium in following embodiment is 200g peeling potatoes, is cut into small pieces, adds water in pot 1000mL boils half an hour, filters with double gauze, takes its filtrate and adds 20g glucose, and adds water and supply 1000mL.
Embodiment 1, the separation of bacterial strain and qualification
One, the separation of bacterial strain W11025 and qualification
1, the separation of bacterial strain W11025
Sample (in October, the 2014 Inner Mongol Wuyuan County) 10.0g of compost will be picked up from, add the nothing of the 90mL of band bead In bacterium water, standing 20min, on rotary shaker, 200r/min fully vibrates 30min, i.e. obtains 1 × 101Dilution bacterium is hanged Liquid, draws the above-mentioned bacteria suspension of 5.0mL with Sterile pipette and joins in the triangular flask equipped with 45mL sterilized water, and fully vibration is shaken Even, obtain 1 × 102Dilution bacteria suspension, method makes 1 × 10 according to this3、1×104、1×105Dilution bacteria suspension.
Aseptically, above each dilution factor bacteria suspension respectively takes 100 μ L and is coated on beef-protein medium, and 20 DEG C cultivate 24-72h, observe and produce with or without bacterium colony, select a strain growth fast, the different bacterial strain of form, and by this Strain Designation For bacterial strain W11025.
2, the qualification of bacterial strain W11025
(1) form and the Physiology and biochemistry of bacterial strain W11025 is identified
The form of bacterial strain W11025 and Physiology and biochemistry qualification result are as shown in Figure 3.
(2) Molecular Identification of bacterial strain W11025
The STb gene extracting bacterial strain W11025 carries out PCR amplification as template, employing universal primer 27f and 1492r, obtains Fragment containing bacterial strain W11025 16S rDNA conserved region.Primer sequence is as follows:
27f (forward primer): 5 '-AGAGTTTGATCCTGGCTCAG-3 '；
1492r (downstream primer): 5 '-TACGGTTACCTTGTTACGACTT-3 '.
The sequence of the 16s rDNA of the bacterial strain W11025 that PCR amplification obtains is as shown in Figure 4.
3, the preservation of bacterial strain W11025
According to morphological characteristic, Physiology and biochemistry and Molecular Identification result and analysis, the Classification And Nomenclature of bacterial strain W11025 is hay Bacillus subtilis, this bacterial strain was preserved in Chinese microorganism strain preservation management on 06 22nd, 2016 CGMCC, address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, the Chinese Academy of Sciences is micro-(are called for short in committee's common micro-organisms center Biological study institute, postcode 100101), deposit number is CGMCC No.12705.
Two, the separation of bacterial strain W160617 and qualification
1, the separation of bacterial strain W160617
Sample (in October, the 2014 Inner Mongol Wuyuan County) 10.0g of compost will be picked up from, add the nothing of the 90mL of band bead In bacterium water, standing 20min, on rotary shaker, 200r/min fully vibrates 30min, i.e. obtains 1 × 101Dilution bacterium is hanged Liquid, draws the above-mentioned bacteria suspension of 5.0mL with Sterile pipette and joins in the triangular flask equipped with 45mL sterilized water, and fully vibration is shaken Even, obtain 1 × 102Dilution bacteria suspension, method makes 1 × 10 according to this3、1×104、1×105Dilution bacteria suspension.
Aseptically, above each dilution factor bacteria suspension respectively takes 100 μ L and is coated on beef-protein medium, and 20 DEG C cultivate 24-72h, observe and produce with or without bacterium colony, select a strain growth fast, the different bacterial strain of form, and by this Strain Designation For bacterial strain W160617.
2, the qualification of bacterial strain W160617
(1) form and the Physiology and biochemistry of bacterial strain W160617 is identified
Shown in the cellular morphology of bacterial strain W160617 and Physiology and biochemistry qualification result Fig. 5.
(2) Molecular Identification of bacterial strain W160617
The STb gene extracting bacterial strain W160617 carries out PCR amplification as template, employing universal primer 27f and 1492r, obtains Fragment containing bacterial strain W160617 16S rDNA conserved region.Primer sequence is as follows:
27f (forward primer): 5 '-AGAGTTTGATCCTGGCTCAG-3 '；
1492r (downstream primer): 5 '-TACGGTTACCTTGTTACGACTT-3 '.
The sequence of the 16s rDNA of the bacterial strain W11025 that PCR amplification obtains is as shown in Figure 6.
3, the preservation of bacterial strain W160617
According to morphological characteristic, Physiology and biochemistry and Molecular Identification result and analysis, the Classification And Nomenclature of bacterial strain W160617 is Xie Dian Afnyloliquefaciens Bacillus amyloliquefaciens, this bacterial strain was preserved in China Microbiological bacterium on 06 22nd, 2016 Kind preservation administration committee common micro-organisms center (abbreviation CGMCC, address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, in Institute of microbiology of academy of science of state, postcode 100101), deposit number is CGMCC No.12703.
Three, the separation of bacterial strain W052501 and qualification
1, the separation of bacterial strain W052501
Sample (in December, the 2015 Pinggu district) 10.0g of compost will be picked up from, add the nothing of the 90mL of band bead In bacterium water, standing 20min, on rotary shaker, 200r/min fully vibrates 30min, i.e. obtains 1 × 101Dilution bacterium is hanged Liquid, draws the above-mentioned bacteria suspension of 5.0mL with Sterile pipette and joins in the triangular flask equipped with 45mL sterilized water, and fully vibration is shaken Even, obtain 1 × 102Dilution bacteria suspension, method makes 1 × 10 according to this3、1×104、1×105Dilution bacteria suspension.
Aseptically, above each dilution factor bacteria suspension respectively takes 100 μ L and is coated in YPD culture medium, cultivates 24-for 20 DEG C 72h, observes and produces with or without bacterium colony, selects a strain growth fast, the different bacterial strain of form, and is bacterial strain by this Strain Designation W052501。
2, the qualification of bacterial strain W052501
(1) identification of morphology of bacterial strain W052501
In YPD culture medium 25 DEG C cultivate three days, cell is avette, oval, sausage shape, size be (3.2-6.0) × (4.9-11.6)μm.YPD medium slant 25 DEG C is cultivated one month, bacterium colony cheese shape, milky, and surface smooths, non-reflective, limit Edge tree root shape.Corn meal agar Dalmau flat board is cultivated, and produces pseudohypha.
(2) Physiology and biochemistry of bacterial strain W052501 is identified
The physicochemical characteristics of bacterial strain W052501 is as shown in Figure 7.
(3) Molecular Identification of bacterial strain W052501
Extract the STb gene of bacterial strain W052501 as template, employing NL1:5'-GCATATCAATAAGCGGAGGAAAAG-3' Carry out PCR amplification with NL4:5'-GGTCCGTGTTTCAAGACGG-3' primer, obtain the rRNA gene D1D2 of bacterial strain W052501 Region sequence, and it is checked order.Sequencing result is as shown in Figure 8.
3, the preservation of bacterial strain
According to morphological characteristic, Physiology and biochemistry and Molecular Identification result and analysis, the Classification And Nomenclature of bacterial strain W052501 is Fructus Hordei Germinatus Sugar candida mycoderma Candida maltosa, this bacterial strain was preserved in Chinese microorganism strain preservation management on 06 22nd, 2016 CGMCC, address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, the Chinese Academy of Sciences is micro-(are called for short in committee's common micro-organisms center Biological study institute, postcode 100101), deposit number is CGMCC No.12702.
Four, the separation of bacterial strain W052502 and qualification
1, the separation of bacterial strain W052502
Sample (in December, the 2014 Pinggu district) 10.0g of compost will be picked up from, add the nothing of the 90mL of band bead In bacterium water, standing 20min, on rotary shaker, 200r/min fully vibrates 30min, i.e. obtains 1 × 101Dilution bacterium is hanged Liquid, draws the above-mentioned bacteria suspension of 5.0mL with Sterile pipette and joins in the triangular flask equipped with 45mL sterilized water, and fully vibration is shaken Even, obtain 1 × 102Dilution bacteria suspension, method makes 1 × 10 according to this3、1×104、1×105Dilution bacteria suspension.
Aseptically, above each dilution factor bacteria suspension respectively takes 100 μ L and is coated in YPD culture medium, cultivates 24-for 20 DEG C 72h, observes and produces with or without bacterium colony, selects a strain growth fast, the different bacterial strain of form, and is bacterial strain by this Strain Designation W052502。
2, the qualification of bacterial strain W052502
(1) identification of morphology of bacterial strain W052502
YPD solid medium 28 DEG C is cultivated 3 days, and bacterium colony is milky white, and without pseudohypha, water logging sheet basis of microscopic observation brings out more Bud, cell is round, and liquid YPD cultivates and produces conveying white precipitate.
(2) Physiology and biochemistry of bacterial strain W052502 is identified
Bacterial strain W052502 Physiology and biochemistry qualification result is as shown in table 1.
Table 1, bacterial strain W052502 Physiology and biochemistry are identified
(3) Molecular Identification of bacterial strain W052502
The STb gene of extraction bacterial strain W052502, as template, uses following primer NL1:5'- GCATATCAATAAGCGGAGGAAAAG-3' and NL4:5'-GGTCCGTGTTTCAAGACGG-3' carries out PCR amplification, obtains bacterium The 26SrDNA gene D1D2 region sequence of strain W052502, sequencing result is as shown in sequence 1 in sequence table.
3, the preservation of bacterial strain W052502
According to morphological characteristic, Physiology and biochemistry and Molecular Identification result and analysis, the Classification And Nomenclature of bacterial strain W052502 is wine brewing Yeast Saccharomyces cerevisiae, this bacterial strain was preserved in Chinese microorganism strain preservation pipe on 06 22nd, 2016 CGMCC, address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, the Chinese Academy of Sciences (are called for short in reason committee's common micro-organisms center Institute of microbiology, postcode 100101), deposit number is CGMCC No.12701.
Five, the separation of bacterial strain W30117 and qualification
1, the separation of bacterial strain W30117
Sample (in October, the 2015 Pinggu district) 10.0g of compost will be picked up from, add the nothing of the 90mL of band bead In bacterium water, standing 20min, on rotary shaker, 200r/min fully vibrates 30min, i.e. obtains 1 × 101Dilution bacterium is hanged Liquid, draws the above-mentioned bacteria suspension of 5.0mL with Sterile pipette and joins in the triangular flask equipped with 45mL sterilized water, and fully vibration is shaken Even, obtain 1 × 102Dilution bacteria suspension, method makes 1 × 10 according to this3、1×104、1×105Dilution bacteria suspension.
Aseptically, above each dilution factor bacteria suspension respectively takes 100 μ L and is coated in PDA culture medium, cultivates 24-for 20 DEG C 72h, observes and produces with or without bacterium colony, selects a strain growth fast, the different bacterial strain of form, and is bacterial strain by this Strain Designation W30117。
2, the qualification of bacterial strain W30117
(1) identification of morphology of bacterial strain W30117
In PDA culture medium, being white, fade to yellow at the beginning of bacterium colony, the later stage is black, and the colourless central authorities of back periphery are slightly Yellowish-brown.Through cultivating 48h on microscope slide, mycelium has separation, and microscope observes counties and cities: conidiophore is born in podocyte On, diameter 15-20pm, be about 2mm, wall thickness and smooth, dome capsule is formed on top, comprehensively one layer of metulae of covering and one layer it on Stigma, long on stigma have bunchiness memnonious spherical, diameter 3.0-4.0 μm；Conidial head is spherical, diameter 700-800 μm.
(2) Physiology and biochemistry of bacterial strain W30117 is identified
Nitrogen source utilizes and test result indicate that: bacterial strain W30117 can be with sodium nitrate, carbamide, ammonium sulfate, bean cake, casein for nitrogen source Growth, under equal conditions, the ability utilizing casein is the strongest, and colony growth is the fastest.The most suitable growth PH of this bacterial strain is 6.0, the suitableeest Growth temperature is 37 DEG C.
(3) Molecular Identification of bacterial strain W30117
The STb gene extracting bacterial strain W30117 carries out PCR amplification as template, employing universal primer ITS1 and ITS4, obtains Containing bacterial strain W052501ITS sequence.Primer sequence is as follows:
ITS1:5′-T CCGTAGGTGAACCTGCGG-3′
ITS4:5′-TCCTC CGCTTATTGATATGC-3′
The sequence of the ITS sequence of the bacterial strain W30117 that PCR amplification obtains is as shown in Figure 9.
3, the preservation of bacterial strain W30117
According to morphological characteristic, Physiology and biochemistry and Molecular Identification result and analysis, the Classification And Nomenclature of bacterial strain W30117 is black fermented preparation Mould Aspergillus niger, this bacterial strain was preserved in China Committee for Culture Collection of Microorganisms on 06 22nd, 2016 CGMCC, address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Chinese Academy of Sciences's microorganism is ground (are called for short in common micro-organisms center Study carefully institute, postcode 100101), deposit number is CGMCC No.12625.
Six, the separation of bacterial strain W060406 and qualification
1, the separation of bacterial strain W060406
Sample (in October, the 2015 Pinggu district) 10.0g of compost will be picked up from, add the nothing of the 90mL of band bead In bacterium water, standing 20min, on rotary shaker, 200r/min fully vibrates 30min, i.e. obtains 1 × 101Dilution bacterium is hanged Liquid, draws the above-mentioned bacteria suspension of 5.0mL with Sterile pipette and joins in the triangular flask equipped with 45mL sterilized water, and fully vibration is shaken Even, obtain 1 × 102Dilution bacteria suspension, method makes 1 × 10 according to this3、1×104、1×105Dilution bacteria suspension.
Aseptically, above each dilution factor bacteria suspension respectively takes 100 μ L and is coated in PDA culture medium, cultivates 24-for 20 DEG C 72h, observes and produces with or without bacterium colony, selects a strain growth fast, the different bacterial strain of form, and is bacterial strain by this Strain Designation W30117。
2, the qualification of bacterial strain W060406
(1) identification of morphology of bacterial strain W060406
After bacterial strain W060406 Cha Shi cultivates 4d, bacterium colony circle, yellow, size about 5.2cm, quality are loose, dry tack free has Concavo-convex, in granular；Within 12nd day, bacterium colony is paved with whole flat board, and color becomes yellow green, and edge is breen.Mycelia has barrier film, Conidiophore length 1.8-2.1mm, diameter 12.1-14.83 μm, coarse have pit, and top capsule enlarges into spherical vesicle, and top capsule is straight Footpath 48.9-51.2 μm, stigma is double-deck, and conidium is concatenated, and conidium is spherical, size 4.3-5.2 μm.
(2) Molecular Identification of bacterial strain W060406
The STb gene extracting bacterial strain W060406 carries out PCR amplification as template, employing universal primer ITS1 and ITS4, obtains Containing bacterial strain W060406ITS sequence.Primer sequence is as follows:
ITS1:5′-T CCGTAGGTGAACCTGCGG-3′
ITS4:5′-TCCTC CGCTTATTGATATGC-3′
The sequence of the ITS sequence of the bacterial strain W060406 that PCR amplification obtains is as shown in sequence 2 in sequence table.
3, the preservation of bacterial strain W060406
According to morphological characteristic, Physiology and biochemistry and Molecular Identification result and analysis, the Classification And Nomenclature of bacterial strain W060406 is rice-koji Mould Aspergillus oryzae, this bacterial strain was preserved in China Committee for Culture Collection of Microorganisms on 06 22nd, 2016 CGMCC, address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Chinese Academy of Sciences's microorganism is ground (are called for short in common micro-organisms center Study carefully institute, postcode 100101), deposit number is CGMCC No.12627.
Seven, the qualification of bacterial strain W2441
1, the separation of bacterial strain W2441
To pick up from sample (in July, the 2015 Daxing district, Beijing, China) 10.0g of compost, the 90mL's of addition band bead is aseptic In water, standing 20min, on rotary shaker, 200r/min fully vibrates 30min, i.e. obtains 1 × 101Dilution bacterium is hanged Liquid, draws the above-mentioned bacteria suspension of 5.0mL with Sterile pipette and joins in the triangular flask equipped with 45mL sterilized water, and fully vibration is shaken Even, obtain 1 × 102Dilution bacteria suspension, method makes 1 × 10 according to this3、1×104、1×105Dilution bacteria suspension.
Aseptically, above each dilution factor bacteria suspension respectively takes 100 μ L and is coated in PDA culture medium, cultivates 24-for 20 DEG C 72h, observes and produces with or without bacterium colony, selects growth fast, the different bacterial strain of form, and is bacterial strain W2441 by this Strain Designation.
2, the qualification of bacterial strain W2441
(1) identification of morphology of bacterial strain W2441
Bacterial strain W2441PDA plating medium Initial stage of culture white mycelium, starts to produce spore after cultivating 60 days, and bacterium colony is in secretly Yellow.Microscope observes mycelia to be had every, Cong Zhizhuan；Conidium is oval or pyriform, directly raw from mycelia side and top.
(2) Molecular Identification of bacterial strain W2441
The STb gene extracting bacterial strain W2441 carries out PCR amplification as template, employing universal primer ITS4 and ITS 5, obtains Containing bacterial strain W2441ITS fragment.Primer sequence is as follows:
ITS4:5’-TCCGTAGGTGAACCTGCGG-3’
ITS5:5’-GCATATCAATAAGCGGAGGA-3’
The ITS sequence of the bacterial strain W2441 that PCR amplification obtains is as shown in sequence 3 in sequence table.
3, the preservation of bacterial strain W2441
According to morphological characteristic, Physiology and biochemistry and Molecular Identification result and analysis, the Classification And Nomenclature of bacterial strain W2441 is thermophilic side Spore mould Sporotrichum thermophile, this bacterial strain was preserved in Chinese microorganism strain preservation pipe on 06 22nd, 2016 CGMCC, address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, the Chinese Academy of Sciences (are called for short in reason committee's common micro-organisms center Institute of microbiology, postcode 100101), deposit number is CGMCC No.12626.
Eight, the separation of bacterial strain W10208 and qualification
1, the separation of bacterial strain W10208
To pick up from sample (in May, the 2015 Pinggu district) 10.0g of compost, the 90mL's of addition band bead is aseptic In water, standing 20min, on rotary shaker, 200r/min fully vibrates 30min, i.e. obtains 1 × 101Dilution bacterium is hanged Liquid, draws the above-mentioned bacteria suspension of 5.0mL with Sterile pipette and joins in the triangular flask equipped with 45mL sterilized water, and fully vibration is shaken Even, obtain 1 × 102Dilution bacteria suspension, method makes 1 × 10 according to this3、1×104、1×105Dilution bacteria suspension.
Aseptically, above each dilution factor bacteria suspension respectively takes 100 μ L and is coated on beef-protein medium, and 20 DEG C cultivate 24-72h, observe and produce with or without bacterium colony, select a strain growth fast, the different bacterial strain of form, and by this Strain Designation For bacterial strain W10208.
2, the qualification of bacterial strain W10208
(1) form and the Physiology and biochemistry of bacterial strain W10208 is identified
Bacterial strain W10208 is colony diameter 0.5-1.5cm on Carnis Bovis seu Bubali cream culture machine, white, sticky, transparent, edge projection, Irregularly, there is gauffer on surface.Physiology and biochemistry qualification result is as shown in table 2.
Table 2, the Physiology and biochemistry qualification result of bacterial strain W10208
(2) Molecular Identification of bacterial strain W10208
The STb gene extracting bacterial strain W10208 carries out PCR amplification as template, employing universal primer 27f and 1492r, obtains Fragment containing bacterial strain W10208 16S rDNA conserved region.Primer sequence is as follows:
27f (forward primer): 5 '-AGAGTTTGATCCTGGCTCAG-3 '；
1492r (downstream primer): 5 '-TACGGTTACCTTGTTACGACTT-3 '.
The sequence of the 16s rDNA of the bacterial strain W10208 that PCR amplification obtains is as shown in sequence 4 in sequence table.
3, the preservation of bacterial strain W10208
According to morphological characteristic, Physiology and biochemistry and Molecular Identification result and analysis, the Classification And Nomenclature of bacterial strain W10208 is thermophilic Bacillus stearothermophilus Bacillus stearothermophilus, this bacterial strain was preserved in Chinese micro-life on 06 22nd, 2016 CGMCC, address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 (are called for short in thing culture presevation administration committee's common micro-organisms center Number, Institute of Microorganism, Academia Sinica, postcode 100101), deposit number is CGMCC No.12704.
Embodiment 2, the preparation of low temperature resistant compost fermentation agent and application thereof
One, the preparation of low temperature resistant compost fermentation agent
1, complex micro organism fungicide is prepared
Take each microbial bacterial agent according to following weight to mix, obtain complex micro organism fungicide: saccharomyces cerevisiae microbial inoculum 1.4kg, Candida maltosa bacteria agent 1.2kg, bacillus subtilis microbial agent 1.0kg, bacillus amyloliquefaciens microbial inoculum 1.4kg, aspergillus oryzae microbial inoculum 1.5kg, aspergillus niger microbial inoculum 1.3kg, bacstearothermophilus microbial inoculum 1.0kg, sporotrichum thermophile bacterium Agent 1.0kg.
The preparation method of above-mentioned Saccharomyces Cerevisiae in S accharomyces cerevisiae microbial inoculum is as follows: by saccharomyces cerevisiae Saccharomyces cerevisiae W052502 bacterial strain (preserving number is CGMCC No.12701) is inoculated in YPD culture medium In, at 29 DEG C, under conditions of 200rmp cultivate 20 hours, then after 3000r/min centrifugal concentrating 20min, with zeolite powder according to Mass ratio is the ratio absorption mixing of 1:7, obtains Saccharomyces Cerevisiae in S accharomyces cerevisiae microbial inoculum.Wherein, wine brewing The concentration of yeast Saccharomyces cerevisiae W052502 bacterial strain is 5,000,000,000 cfu/g.
The preparation method of above-mentioned Candida maltosa Candida maltosa microbial inoculum is: by Candida maltosa Candida maltosa W052501 bacterial strain (preserving number is CGMCC No.12702) is inoculated in YPD culture medium, at 30 DEG C, Cultivate 22 hours under the conditions of 200rmp, then after 3000r/min centrifugal concentrating 20min, be 1:7's with zeolite powder according to mass ratio Ratio absorption mixing, obtains Candida maltosa Candida maltosa microbial inoculum.Wherein, Candida maltosa Candida The concentration of maltosa W052501 bacterial strain is 6,000,000,000 cfu/g.
The preparation method of above-mentioned bacillus subtilis Bacillus subtilis microbial inoculum is: by bacillus subtilis In Bacillus subtilis W11025 bacterial strain (preserving number is CGMCC No.12705) inoculation beef-protein medium, At 30 DEG C, cultivate under the conditions of 230rmp to spore forming rate and reach 85%, then after 3000r/min centrifugal concentrating 20min, with zeolite Powder, according to the ratio absorption mixing that mass ratio is 1:5, obtains bacillus subtilis Bacillus subtilis microbial inoculum.Wherein, withered The concentration of grass bacillus subtilis W11025 bacterial strain is 50,000,000,000 cfu/g.
The preparation method of above-mentioned bacillus amyloliquefaciens Bacillus amyloliquefaciens microbial inoculum is: will solve starch Bacillus amyloliquefaciens W160617 bacterial strain (preserving number is CGMCC No.12703) is inoculated in In beef-protein medium, at 31 DEG C, cultivate under the conditions of 230rmp to spore forming rate and reach 85%, then 3000r/min After centrifugal concentrating 20min, with zeolite powder according to the ratio absorption mixing that mass ratio is 1:5, obtain bacillus amyloliquefaciens Bacillus amyloliquefaciens microbial inoculum.Wherein, bacillus amyloliquefaciens Bacillus amyloliquefaciens The concentration of W160617 bacterial strain is 50,000,000,000 cfu/g.
The preparation method of above-mentioned aspergillus oryzae Aspergillus oryzae microbial inoculum is: by aspergillus oryzae Aspergillus Oryzae W060406 bacterial strain (preserving number is CGMCC No.12627) is inoculated in PDA culture medium, at 28 DEG C, and 100rmp condition Lower cultivation 18h, then after 3000r/min centrifugal concentrating 20min, with zeolite powder according to the ratio absorption mixing that mass ratio is 1:5, Obtain aspergillus oryzae Aspergillus oryzae microbial inoculum.Wherein, aspergillus oryzae Aspergillus oryzae W060406 bacterial strain Concentration is 5,000,000,000 cfu/g.
The preparation method of above-mentioned aspergillus niger Aspergillus niger microbial inoculum is: by aspergillus niger Aspergillus niger W30117 bacterial strain (preserving number is CGMCC No.12625) is inoculated in PDA culture medium, at 32 DEG C, cultivates under the conditions of 100rmp 20h, then after 3000r/min centrifugal concentrating 20min, with zeolite powder according to the ratio absorption mixing that mass ratio is 1:5, obtains black Aspergillosis Aspergillus niger microbial inoculum.Wherein, the concentration of aspergillus niger Aspergillus niger W30117 bacterial strain is 3,000,000,000 cfu/g。
The preparation method of above-mentioned bacstearothermophilus Bacillus stearothermophilus microbial inoculum is: will be addicted to Hot Bacillus stearothermophilus Bacillus stearothermophilus W10208 bacterial strain (preserving number is CGMCC No.12704) It is inoculated in beef-protein medium, at 40 DEG C, cultivates under the conditions of 230rmp to spore forming rate and reach 85%, then After 3000r/min centrifugal concentrating 20min, with zeolite powder according to the ratio absorption mixing that mass ratio is 1:5, obtain stearothermophilus bud Spore bacillus Bacillus stearothermophilus microbial inoculum.Wherein, bacstearothermophilus Bacillus The concentration of stearothermophilus W10208 bacterial strain is 10,000,000,000 cfu/g.
The preparation method of above-mentioned sporotrichum thermophile Sporotrichum thermophile microbial inoculum is: by sporotrichum thermophile Sporotrichum thermophile W2441 bacterial strain (preserving number is CGMCC No.12626) is inoculated in PDA culture medium, At 45 DEG C, cultivate 20h under the conditions of 100rmp, then after 3000r/min centrifugal concentrating 20min, with zeolite powder according to mass ratio be The ratio absorption mixing of 1:5, obtains sporotrichum thermophile Sporotrichum thermophile microbial inoculum.Wherein, sporotrichum thermophile The concentration of Sporotrichum thermophile W2441 bacterial strain is 2,000,000,000 cfu/g.
Above-described embodiment relates to strain and each strain growth characteristics under cryogenic are as shown in table 3.
Table 3, above-described embodiment relate to strain and each strain growth characteristics under cryogenic
2, low temperature resistant compost fermentation agent is prepared
Weigh the complex micro organism fungicide 9.8kg that step 1 obtains, and (flourishing age is thrown by state with 90.2kg zeolite powder carrier by it Chengde Science and Technology Ltd.) mixing, obtain the decomposing agent of the present invention, be low temperature resistant compost fermentation agent.
Two, low temperature resistant compost fermentation agent application in fermentation fertilizer
Test site is Pinggu district's Tai Fengping Organic Fertilizer Plants, has with chicken manure, mushroom residue mixture for fermenting raw materials Machine fertilizer ferments, fertilizer after being fermented.Test period is January 4.
Test carries out fermentation according to whether interpolation decomposing agent in fermenting raw materials fertilizer and is divided into following two groups:
1, low temperature resistant compost fermentation agent processes (decomposing agent process group): every 1000kg fermenting raw materials fertilizer adds above-mentioned step The low temperature resistant compost fermentation agent 5kg of a rapid preparation；
2, without decomposing agent space management: without decomposing agent in fermenting raw materials fertilizer.
The online monitor of GMS100 using Beijing Taihua Hengyue Technology Development Co., Ltd. to produce continues to monitor Heap temperature change and heap body H in sweat2S、SO2、NO2、NH3Discharge capacity.Fermentation heap temperature situation of change is shown in Fig. 1, useless Gas discharge capacity the results are shown in Table 4.
Table 4, discharge amount of exhaust gas
Hydrogen sulfide (ppm) Sulfur dioxide (ppm) Nitrogen dioxide (ppm) Ammonia (ppm)
Decomposing agent processes 50803.5 4495.1 137.9 1675804
Space management 42083.2 34625.1 5300.1 2159965
Slip (%) 17.16 87.02 97.40 22.42
As can be seen from the above results, after using the decomposing agent of the present invention to process, relative to blank, fermentation heap body liter Temperature speed accelerates, and hydrogen sulfide, sulfur dioxide, nitrogen dioxide, the discharge capacity of 4 kinds of waste gas of ammonia all reduce, and wherein hydrogen sulfide subtracts Row leads and reaches 17.16%, and sulfur dioxide reduction of discharging rate reaches 87.02%, and nitrogen dioxide reduction of discharging rate reaches 97.4%, and ammonia reduction of discharging rate reaches 22.42%.
Embodiment 3, the preparation of low temperature resistant compost fermentation agent and application thereof
One, the preparation of low temperature resistant compost fermentation agent
1, complex micro organism fungicide is prepared
Take each microbial bacterial agent according to following weight to mix, obtain complex micro organism fungicide: saccharomyces cerevisiae microbial inoculum 1.2kg, Candida maltosa bacteria agent 1.3kg, bacillus subtilis microbial agent 0.8kg, bacillus amyloliquefaciens microbial inoculum 1.4kg, aspergillus oryzae microbial inoculum 1.5kg, aspergillus niger microbial inoculum 1.3kg, bacstearothermophilus microbial inoculum 0.8kg, sporotrichum thermophile bacterium Agent 0.8kg.The most respective living bacteria count of each bacteria agent is respectively as follows: saccharomyces cerevisiae 5,000,000,000 cfu/g, maltose vacation silk ferment Female 5,000,000,000 cfu/g, bacillus subtilis 30,000,000,000 cfu/g, bacillus amyloliquefaciens 50,000,000,000 cfu/g, aspergillus oryzae 2,000,000,000 cfu/g, Aspergillus niger 2,000,000,000 cfu/g, bacstearothermophilus 20,000,000,000 cfu/g, sporotrichum thermophile 2,000,000,000 cfu/g.
The preparation method of above-mentioned Saccharomyces Cerevisiae in S accharomyces cerevisiae microbial inoculum is as follows: by saccharomyces cerevisiae Saccharomyces cerevisiae W052502 bacterial strain (preserving number is CGMCC No.12701) is inoculated in YPD culture medium In, at 29 DEG C, under conditions of 200rmp cultivate 20 hours, then after 3000r/min centrifugal concentrating 20min, with zeolite powder according to Mass ratio is the ratio absorption mixing of 1:7, obtains Saccharomyces Cerevisiae in S accharomyces cerevisiae microbial inoculum.Wherein, wine brewing The concentration of yeast Saccharomyces cerevisiae W052502 bacterial strain is 5,000,000,000 cfu/g.
The preparation method of above-mentioned Candida maltosa Candida maltosa microbial inoculum is: by Candida maltosa Candida maltosa W052501 bacterial strain (preserving number is CGMCC No.12702) is inoculated in YPD culture medium, at 30 DEG C, Cultivate 22 hours under the conditions of 200rmp, then after 3000r/min centrifugal concentrating 20min, be 1:7's with zeolite powder according to mass ratio Ratio absorption mixing, obtains Candida maltosa Candida maltosa microbial inoculum.Wherein, Candida maltosa Candida The concentration of maltosa W052501 bacterial strain is 6,000,000,000 cfu/g.
The preparation method of above-mentioned bacillus subtilis Bacillus subtilis microbial inoculum is: by bacillus subtilis In Bacillus subtilis W11025 bacterial strain (preserving number is CGMCC No.12705) inoculation beef-protein medium, At 30 DEG C, cultivate under the conditions of 230rmp to spore forming rate and reach 85%, then after 3000r/min centrifugal concentrating 20min, with zeolite Powder, according to the ratio absorption mixing that mass ratio is 1:5, obtains bacillus subtilis Bacillus subtilis microbial inoculum.Wherein, withered The concentration of grass bacillus subtilis W11025 bacterial strain is 50,000,000,000 cfu/g.
The preparation method of above-mentioned bacillus amyloliquefaciens Bacillus amyloliquefaciens microbial inoculum is: will solve starch Bacillus amyloliquefaciens W160617 bacterial strain (preserving number is CGMCC No.12703) is inoculated in In beef-protein medium, at 31 DEG C, cultivate under the conditions of 230rmp to spore forming rate and reach 85%, then 3000r/min After centrifugal concentrating 20min, with zeolite powder according to the ratio absorption mixing that mass ratio is 1:5, obtain bacillus amyloliquefaciens Bacillus amyloliquefaciens microbial inoculum.Wherein, bacillus amyloliquefaciens Bacillus amyloliquefaciens The concentration of W160617 bacterial strain is 50,000,000,000 cfu/g.
The preparation method of above-mentioned aspergillus oryzae Aspergillus oryzae microbial inoculum is: by aspergillus oryzae Aspergillus Oryzae W060406 bacterial strain (preserving number is CGMCC No.12627) is inoculated in PDA culture medium, at 28 DEG C, and 100rmp condition Lower cultivation 18h, then after 3000r/min centrifugal concentrating 20min, with zeolite powder according to the ratio absorption mixing that mass ratio is 1:5, Obtain aspergillus oryzae Aspergillus oryzae microbial inoculum.Wherein, aspergillus oryzae Aspergillus oryzae W060406 bacterial strain Concentration is 5,000,000,000 cfu/g.
The preparation method of above-mentioned aspergillus niger Aspergillus niger microbial inoculum is: by aspergillus niger Aspergillus niger W30117 bacterial strain (preserving number is CGMCC No.12625) is inoculated in PDA culture medium, at 32 DEG C, cultivates under the conditions of 100rmp 20h, then after 3000r/min centrifugal concentrating 20min, with zeolite powder according to the ratio absorption mixing that mass ratio is 1:5, obtains black Aspergillosis Aspergillus niger microbial inoculum.Wherein, the concentration of aspergillus niger Aspergillus niger W30117 bacterial strain is 3,000,000,000 cfu/g。
The preparation method of above-mentioned bacstearothermophilus Bacillus stearothermophilus microbial inoculum is: will be addicted to Hot Bacillus stearothermophilus Bacillus stearothermophilus W10208 bacterial strain (preserving number is CGMCC No.12704) It is inoculated in beef-protein medium, at 40 DEG C, cultivates under the conditions of 230rmp to spore forming rate and reach 85%, then After 3000r/min centrifugal concentrating 20min, with zeolite powder according to the ratio absorption mixing that mass ratio is 1:5, obtain stearothermophilus bud Spore bacillus Bacillus stearothermophilus microbial inoculum.Wherein, bacstearothermophilus Bacillus The concentration of stearothermophilus W10208 bacterial strain is 10,000,000,000 cfu/g.
The preparation method of above-mentioned sporotrichum thermophile Sporotrichum thermophile microbial inoculum is: by sporotrichum thermophile Sporotrichum thermophile W2441 bacterial strain (preserving number is CGMCC No.12626) is inoculated in PDA culture medium, At 45 DEG C, cultivate 20h under the conditions of 100rmp, then after 3000r/min centrifugal concentrating 20min, with zeolite powder according to mass ratio be The ratio absorption mixing of 1:5, obtains sporotrichum thermophile Sporotrichum thermophile microbial inoculum.Wherein, sporotrichum thermophile The concentration of Sporotrichum thermophile W2441 bacterial strain is 2,000,000,000 cfu/g.
2, low temperature resistant compost fermentation agent is prepared
Weigh the complex micro organism fungicide 9.1kg that step 1 obtains, and (flourishing age Chengde is thrown by state with 90.9kg zeolite powder by it Science and Technology Ltd.) carrier mixing, obtain the decomposing agent of the present invention, be low temperature resistant compost fermentation agent.
Two, low temperature resistant compost fermentation agent application in fermentation fertilizer
Test site is Pinggu district's Tai Fengping Organic Fertilizer Plants, with chicken manure, Flammulina velutiper (Fr.) Sing slag mixture as fermenting raw materials Fertilizer ferments, fertilizer after being fermented.Test period is March 7.
Test carries out fermentation according to whether interpolation decomposing agent in fermenting raw materials fertilizer and is divided into following two groups:
1, low temperature resistant compost fermentation agent processes (decomposing agent process group): every 1000kg fermenting raw materials fertilizer adds above-mentioned step The low temperature resistant compost fermentation agent 5kg of a rapid preparation；
2, without decomposing agent space management: without decomposing agent in fermenting raw materials fertilizer.
The online monitor of GMS100 using Beijing Taihua Hengyue Technology Development Co., Ltd. to produce continues to monitor Heap temperature change and heap body H in sweat2S、SO2、NO2、NH3Discharge capacity.Fermentation heap temperature situation of change is shown in Fig. 2, useless Gas discharge capacity the results are shown in Table 5.
Table 5, discharge amount of exhaust gas
Hydrogen sulfide (ppm) Sulfur dioxide (ppm) Nitrogen dioxide (ppm) Ammonia (ppm)
Decomposing agent processes 11453.1 2476.9 10.84 926261
Space management 34448.4 21734.5 2465.3 3220152
Slip (%) 66.75 88.60 99.56 71.23
As can be seen from the above results, after using the decomposing agent of the present invention to process, relative to blank, fermentation heap body liter Temperature speed accelerates, and hydrogen sulfide, sulfur dioxide, nitrogen dioxide, the discharge capacity of 4 kinds of waste gas of ammonia all reduce, and wherein hydrogen sulfide subtracts Row leads and reaches 66.75%, and sulfur dioxide reduction of discharging rate reaches 88.60%, and nitrogen dioxide reduction of discharging rate reaches 99.56%, and ammonia reduction of discharging rate reaches 71.23%.
From above experimental result it can be seen that use the decomposing agent of the present invention, relative to blank, it is possible to accelerate to become thoroughly decomposed Process programming rate, is warming up to 50 DEG C (under weather conditions of temperature on average 20 DEG C) for 12 hours in advance, and digest process high temperature Phase temperature is higher, and the cooldown period that becomes thoroughly decomposed occurs fast.It addition, after using the leaven of the present invention to process, relative to blank, compost Process reaches 22.42-71.23% to the ammonia minimizing of environmental emission, and hydrogen sulfide reduces 17.16-66.75%, it is possible to be greatly reduced The concentration of the gas such as ammonia and hydrogen sulfide in environment, reduces the discharge of composting process stink further.
Embodiment 4, compliance test result experiment (using Caulis et Folium Lactucae Sativae) of low temperature resistant compost fermentation agent
The fertilizer produced for compost fermentation agent fermentation poultry dung and the mushroom residue of the checking present invention is the most effective in field, After the fermentation obtain embodiment 2, organic fertilizer application carries out field test on Caulis et Folium Lactucae Sativae, and detects field efficacy.Concrete steps are such as Under:
One, test method
1, test period and place
Test arrangement is carried out at Fangshan, Beijing Lu Cun, and test period is in July, 2015 in March, 2015 to.
2, for examination soil Elemental characters
It is shown in Table 6 for examination soil basic physicochemical character measurement result.
Table 6, testing site soil nutrient basic condition
3, for studying thing
It is Caulis et Folium Lactucae Sativae for studying thing.
4, test process and land for growing field crops are arranged
This test arranges following three and processes, in triplicate, each process community random alignment, plot area 20m2Above.
Process one: after the fermentation that in embodiment 2 prepared by step 2, fertilizer processes, fertilizer 200kg/ after bottom application fermentation 667m2, impose carbamide 10kg/667m2。
Process two: blank (CK), base manure is not executed with topdressing.
Process three: conventional fertilizer application, bottom application compound fertilizer (it is purchased from Beijing Agriculture means of production company limited, total nutrient content >= 25%) 200kg/667m2, impose carbamide 10kg/667m2。
5, field management
(1) field planting with gather
Trial crops is bile ingredients, and kind is emperor.March 12 started nursery, and April 26 used base manure, after intertillage, Ditching, ridging, field planting on April 27, plantation density is 6100 strains/667m2.Process one and be respectively May 20 and June 2 with processing three Day imposes carbamide twice, each consumption 5kg/667m2, use ditch spread, water after earthing.
(2) water and intertill
At Caulis et Folium Lactucae Sativae seedling-slowing stage, trophophase, water in time during balling initial soil arid, after watering, carry out weeding, middle farming Industry.
(3) gather and survey product
July 5, ripe results, recorded each community actual production.
(4) field investigation
6, biological character investigation
Each community selects the representational 15 strain Caulis et Folium Lactucae Sativae investigation biological characters that size is similar with growing way.
7, yield investigation
After fruit maturation, gather respectively by community and weigh, and record yield.
Two, result and analysis
1, the different disposal impact on Caulis et Folium Lactucae Sativae biological character
In the seated phase, each community chooses representational 5 strains respectively, cleans its plant height of " Invest, Then Investigate ", root length, root fresh weight and does Weight.The root of Caulis et Folium Lactucae Sativae being placed in baking oven 105 DEG C complete 15 minutes, 80 DEG C of constant temperature, after 48 hours, claim its weight to be root dry weight.
Result is as shown in table 7.As can be seen from Table 7, test processes one compared with other process: plant height increases 0.4-3.4cm, Root length increases 0.3-1.8cm, and root fresh weight increases 0.5-3.2g, and root dry weight increases 0.2-0.8g.
Table 7, Caulis et Folium Lactucae Sativae fertilizer test respectively process biological character
Process number Plant height cm Root length cm Root fresh weight g Root dry weight g
Process one 24.1 11.3 18.3 2.6
Process two 20.7 9.5 15.1 1.8
Process three 23.7 11.0 17.8 2.4
2, the different fertilization impact on Caulis et Folium Lactucae Sativae yield
Caulis et Folium Lactucae Sativae each community collection period counts product respectively, equivalent 667m2Yield, the results are shown in Table 8.As can be seen from Table 8, test processes One compared with other process, and yield substantially increases.
Table 8, different disposal yield result
To above-mentioned yield result variance analysis, the results are shown in Table 9.The results of analysis of variance shows: difference pole between the process of testing site Significantly, between repetition, difference is the most notable.Further the yield result of table 7 is carried out multiple comparisons, the results are shown in Table 10.Permissible by table 10 Finding out, experiment processes one and increases production compared with processing three significantly, but average product relatively processes three volume increase 38.466kg/667m2；Place Reason one relatively processes two differences and reaches the most notable.
Table 9, the variance analysis of yield result
Source of variation Degree of freedom Quadratic sum Mean square F value F0.05 F0.01
Between district's group 2 3080.44 1540.22 0.26X 6.94 18.0
Between process 2 635032.0 317516.0 53.0 6.94 18.0
Error 4 23963.56 5990.89
Amount to 8 662076.0
Note: difference is extremely notable, and significant difference, x difference is the most notable
Table 10, different disposal yield the result of multiple comparisons
Note: letter identical table differential is not different notable, letter is different represents significant difference.
The above results shows: utilize the compost fermentation agent fermentation poultry dung of the present invention and the fertilizer of mushroom residue production to promote Enter Growth of Lettuce, fertilizer process after Caulis et Folium Lactucae Sativae at plant height, root length, root fresh weight, root dry weight is above conventional fertilizer application and blank is right According to process, yield relatively space management difference reaches pole significant level, more conventional fertilising mu volume increase 38.466kg.Proof utilizes the present invention Compost fermentation agent fermentation poultry dung and mushroom residue produce fertilizer there is certain fertilizer efficiency.
Embodiment 5, compliance test result experiment (using peach tree) of low temperature resistant compost fermentation agent
The fertilizer produced for compost fermentation agent fermentation poultry dung and the mushroom residue of the checking present invention is the most effective in field, After the fermentation obtain embodiment 3, organic fertilizer application carries out field test on peach tree, and detects field efficacy.Concrete steps are such as Under:
One, test method
1, test period and place
Test arrangement Beigong village and Zhen Luoying behind big Huashan town, Pinggu District are carried out, and test period is that in JIUYUE, 2014 arrives In October, 2015.
2, for examination soil Elemental characters
It is shown in Table 11 for examination soil basic physicochemical character measurement result.
Table 11, testing site soil nutrient basic condition
3, for studying thing
Being peach tree for studying thing, variety name is bright red.
4, test process and land for growing field crops are arranged
This test arranges following three and processes, in triplicate, and each process community random alignment.Plot area 111m2, row strain Away from 4*2.5m, planting density 40 strains/667m2, the age of tree is 3 years.
Process one: after the fermentation that in embodiment 3 prepared by step 2, fertilizer processes, fertilizer 330kg/ after bottom application fermentation 667m2, impose Chemical Mixed Fertilizer 50kg/667m2。
Process two: blank (CK), base manure is not executed with topdressing.
Process three: conventional fertilizer application, bottom application compound fertilizer (it is purchased from Beijing Agriculture means of production company limited, total nutrient content >= 25%) 100kg/667m2, impose carbamide 50kg/667m2。
5, field management
(1) fertilization time
Base manure is used in by the end of September, 2014 in, topdresses and uses in by the end of March, 2015 in, in addition to fertilising difference, and other control measures Unanimously.
(2) fertilizing method
Wide 25cm is opened in tree crown periphery, the annular ditch of deep 15cm, base fertilizer or topdress uniformly is applied in ditch, then earthing is placed on Above.
(3) water and intertill
The season of growth waters in good time, carries out intertillage operation after watering in time.
(4) other management
The method using artificial pollination and honeybee pollination to combine improves fruit-setting rate；Flower thinning when blooming, fixed fruit before bagging, typically Each inflorescence stays 1 fruit, stays 2 fruits every 0.15 meter, stays high quality fruit, mid-June bagging.Gather first 25 days de-bags.After de-bag Carry out priming leaf, turn the raising qualities such as fruit.
6, field investigation
(1) biological character investigation
5 strains that each community selects tree body size similar with growing way carry out biological character and fruit quality investigation.
(2) yield investigation
After mid-September fruit maturation, start to gather.Gathering by individual plant, each process is gathered 5 strains, weighs, and by little differentiation Do not record yield.
Two, test results and analysis
1, the different fertilization impact on peach tree biological character
At peach tree trophophase, biological character and fruit quality are carried out investigation to record, the results are shown in Table 12.Permissible from table 12 Finding out, the full-bloom stage of three process in two testing sites, young fruit period are consistent with the period of maturation.The leaf color of process one and process three is relatively Other processes deep.Processing one to increase than other process single fruit weight, sugar content, malformed fruit rate has declined, wherein rear Beigong Testing site, village, processes one and relatively processes two (blank) single fruit weight increase 13.1g, and sugar content increases by 1.5%, and malformed fruit rate reduces 3.6%.Luo Ying village, town processes one and relatively processes two (blank) single fruit weight increase 18.1g, and sugar content increases by 0.9%, malformed fruit Rate reduces by 4.2%.From above comparison it can be seen that application of organic fertilizers can be obviously improved peach tree plant biological character, favorably In peach tree high yield, stable yields.
Table 12, peach tree fertilizer test respectively process biological character and quality survey
2, the different fertilization impact on peach tree yield
Peach tree each community collection period counts product respectively, is converted into 667m2Yield, the results are shown in Table 13.As can be seen from Table 13, Each process group one average productivity of each test site is above process group two and the average productivity of process group three.
Above-mentioned yield result is carried out variance analysis, the results are shown in Table 14.The results of analysis of variance shows: between two testing sites process Significant difference, between repetition, difference is the most notable.Further the yield result of table 12 is carried out multiple comparisons, the results are shown in Table 15.By table 15 it can be seen that testing site, rear Beigong village, and process one is compared with process two, process three, and yield all has to be increased in various degree, respectively Increase 1357.3kg/667m2、324.1kg/667m2, rate of growth is respectively 16.0%, 3.4%, and processes one and process two products Amount reaches pole significant difference；Luo Ying testing site, town, processes one compared with processing two, processing three by increasing production in various degree, yield 1707.0kg/667m respectively2、299.5kg/667m2, rate of growth is respectively 20.6%, 3.1%, wherein processes one and produces with processing two Amount difference reaches significant level.
Table 13, different disposal yield result
Table 14, the variance analysis of yield result
Table 15, different disposal yield the result of multiple comparisons
Above-mentioned two testing site result shows, utilizes the compost fermentation agent fermentation poultry dung of the present invention and having of mushroom residue production Machine fertilizer processes the Fructus Persicae more conventional fertilising of malformed fruit rate and blank all decreases, and single fruit weight and the sugar content of fruit have carried High.Wherein malformed fruit rate reduces 0.7-3.6%, and single fruit weight improves 9.9-25.6g, and the sugar content of fruit improves 0.5-1.5%, yield Improve more than 3.0%.Result shows: the fertilizer utilizing the compost fermentation agent fermentation poultry dung of the present invention and mushroom residue to produce can Reduce peach tree malformed fruit rate, improve single fruit weight, improve fruit quality, improve Fructus Persicae yield.Prove to utilize the compost fermentation of the present invention The fertilizer that agent fermentation poultry dung and mushroom residue produce has certain fertilizer efficiency.

Claims (10)

1. a complex micro organism fungicide, its active component is Saccharomyces Cerevisiae in S accharomyces cerevisiae, maltose Candida mycoderma Candida maltosa, bacillus subtilis Bacillus subtilis, bacillus amyloliquefaciens Bacillus Amyloliquefaciens, aspergillus oryzae Aspergillus oryzae, aspergillus niger Aspergillus niger, stearothermophilus bud Spore bacillus Bacillus stearothermophilus and sporotrichum thermophile Sporotrichum thermophile.
Complex micro organism fungicide the most according to claim 1, it is characterised in that: described Saccharomyces Cerevisiae in S accharomyces Cerevisiae, described Candida maltosa Candida maltosa, described bacillus subtilis Bacillus Subtilis, described bacillus amyloliquefaciens Bacillus amyloliquefaciens, described aspergillus oryzae Aspergillus Oryzae, described aspergillus niger Aspergillus niger, described bacstearothermophilus Bacillus The proportioning of the cfu of stearothermophilus and described sporotrichum thermophile Sporotrichum thermophile is (30- 60): (30-60): 500:(200-500): (20-50): (20-50): (100-200): (20-30)；
Or, described Saccharomyces Cerevisiae in S accharomyces cerevisiae, described Candida maltosa Candida Maltosa, described bacillus subtilis Bacillus subtilis, described bacillus amyloliquefaciens Bacillus Amyloliquefaciens, described aspergillus oryzae Aspergillus oryzae, described aspergillus niger Aspergillus niger, institute State bacstearothermophilus Bacillus stearothermophilus and described sporotrichum thermophile Sporotrichum The proportioning of the cfu of thermophile is specially 5:5:30:50:2:2:20:2 or 5:6:50:50:5:3:10:2；
Or, described Saccharomyces Cerevisiae in S accharomyces cerevisiae is specially Saccharomyces Cerevisiae in S accharomyces cerevisiae W052502CGMCC No.12701；
Described Candida maltosa Candida maltosa is specially Candida maltosa Candida maltosa W052501CGMCC No.12702；
Described bacillus subtilis Bacillus subtilis is specially bacillus subtilis Bacillus subtilis W11025 CGMCC No.12705；
Described bacillus amyloliquefaciens Bacillus amyloliquefaciens is specially bacillus amyloliquefaciens Bacillus amyloliquefaciens W160617CGMCC No.12703；
Described aspergillus oryzae Aspergillus oryzae is specially aspergillus oryzae Aspergillus oryzae W060406 CGMCC No.12627；
Described aspergillus niger Aspergillus niger is specially aspergillus niger Aspergillus niger W30117 CGMCC No.12625；
Described bacstearothermophilus Bacillus stearothermophilus is specially bacstearothermophilus Bacillus stearothermophilus W10208CGMCC No.12704；
Described sporotrichum thermophile Sporotrichum thermophile is specially sporotrichum thermophile Sporotrichum thermophile W2441CGMCC No.12626。
3. a preparation method for complex micro organism fungicide, for by Saccharomyces Cerevisiae in S accharomyces cerevisiae microbial inoculum, Candida maltosa Candida maltosa microbial inoculum, bacillus subtilis Bacillus subtilis microbial inoculum, solution starch bud Spore bacillus Bacillus amyloliquefaciens microbial inoculum, aspergillus oryzae Aspergillus oryzae microbial inoculum, aspergillus niger Aspergillus niger microbial inoculum, bacstearothermophilus Bacillus stearothermophilus microbial inoculum and thermophilic side Spore mould Sporotrichum thermophile microbial inoculum is uniformly mixed so as to obtain.
Method the most according to claim 3, it is characterised in that:
Described Saccharomyces Cerevisiae in S accharomyces cerevisiae microbial inoculum, described Candida maltosa Candida Maltosa microbial inoculum, described bacillus subtilis Bacillus subtilis microbial inoculum, described bacillus amyloliquefaciens Bacillus Amyloliquefaciens microbial inoculum, described aspergillus oryzae Aspergillus oryzae microbial inoculum, described aspergillus niger Aspergillus Niger microbial inoculum, described bacstearothermophilus Bacillus stearothermophilus microbial inoculum and described sporotrichum thermophile The mass ratio of Sporotrichum thermophile microbial inoculum is (1-3): (1-3): 2:(1-3): (1-3): (1-3): (1-2): (1-2)；
Or, described Saccharomyces Cerevisiae in S accharomyces cerevisiae microbial inoculum, described Candida maltosa Candida Maltosa microbial inoculum, described bacillus subtilis Bacillus subtilis microbial inoculum, described bacillus amyloliquefaciens Bacillus Amyloliquefaciens microbial inoculum, described aspergillus oryzae Aspergillus oryzae microbial inoculum, described aspergillus niger Aspergillus Niger microbial inoculum, described bacstearothermophilus Bacillus stearothermophilus microbial inoculum and described sporotrichum thermophile The mass ratio of Sporotrichum thermophile microbial inoculum is specially 12:10:14:12:10:14:15:13 or 12:13:8: 14:15:13:8:8；
Or, described Saccharomyces Cerevisiae in S accharomyces cerevisiae is specially Saccharomyces Cerevisiae in S accharomyces cerevisiae W052502CGMCC No.12701；
Described Candida maltosa Candida maltosa is specially Candida maltosa Candida maltosa W052501CGMCC No.12702；
Described bacillus subtilis Bacillus subtilis is specially bacillus subtilis Bacillus subtilis W11025CGMCC No.12705；
Described bacillus amyloliquefaciens Bacillus amyloliquefaciens is specially bacillus amyloliquefaciens Bacillus amyloliquefaciens W160617CGMCC No.12703；
Described aspergillus oryzae Aspergillus oryzae is specially aspergillus oryzae Aspergillus oryzae W060406 CGMCC No.12627；
Described aspergillus niger Aspergillus niger is specially aspergillus niger Aspergillus niger W30117 CGMCC No.12625；
Described bacstearothermophilus Bacillus stearothermophilus is specially bacstearothermophilus Bacillus stearothermophilus W10208CGMCC No.12704；
Described sporotrichum thermophile Sporotrichum thermophile is specially sporotrichum thermophile Sporotrichum thermophile W2441CGMCC No.12626。
5. complex micro organism fungicide described in claim 1 or 2 or by answering that the method described in claim 3 or 4 prepares Close microbial bacterial agent application in following (1)-(8) in any one:
(1) compost fermentation agent or decomposing agent are prepared；
(2) compost；
(3) organic materials or the fermentation of organic waste；
(4) programming rate of fermentation heap body is accelerated；
(5) discharge of odorous gas in digest process is reduced；
(6) plant height of plant and/or root length and/or root fresh weight and/or root dry weight and/or yield are improved；
(7) the malformed fruit rate of Tree Fruit is reduced；
(8) single fruit weight of Tree Fruit and/or sugar content and/or yield are improved；
Described odorous gas is hydrogen sulfide and/or sulfur dioxide and/or nitrogen dioxide and/or ammonia.
6., for a decomposing agent for compost, it is by the complex micro organism fungicide described in claim 1 or 2 or by claim 3 Or the complex micro organism fungicide that the method described in 4 prepares forms with carrier.
Decomposing agent the most according to claim 6, it is characterised in that: described carrier be powdered rice hulls or zeolite powder or powdered rice hulls with The mixture of zeolite powder；
Or, the fineness of described carrier is more than or equal to 60 mesh；
Or, described complex micro organism fungicide is (49-201) with the mass ratio of described carrier: 909；
Or, described complex micro organism fungicide is specially 49:451 or 91:909 with the mass ratio of described carrier.
8. the decomposing agent described in claim 6 or 7 is following 1)-7) in application in any one:
1) compost；
2) organic materials or the fermentation of organic waste；
3) programming rate of fermentation heap body is accelerated；
4) discharge of odorous gas in digest process is reduced；
5) plant height of plant and/or root length and/or root fresh weight and/or root dry weight and/or yield are improved；
6) the malformed fruit rate of Tree Fruit is reduced；
7) single fruit weight of Tree Fruit and/or sugar content and/or yield are improved；
Described odorous gas is hydrogen sulfide and/or sulfur dioxide and/or nitrogen dioxide and/or ammonia.
9. a method for compost, comprises the steps: process organic materials with the decomposing agent described in claim 7 or 8 or have Machine garbage, it is achieved compost；
Or, described decomposing agent is (1-5) with the mass ratio of described organic materials or organic waste: 1000.
10. a preparation method for organic fertilizer, comprises the steps: to process organic with the decomposing agent described in claim 7 or 8 Material or organic waste, obtain organic fertilizer；
Or, the method described in claim 10 organic fertilizer prepared；
Or, the organic fertilizer described in claim 10 is at following a1)-a3) in application in any one:
A1) plant height of plant and/or root length and/or root fresh weight and/or root dry weight and/or yield are improved；
A2) the malformed fruit rate of Tree Fruit is reduced；
A3) single fruit weight of Tree Fruit and/or sugar content and/or yield are improved.
CN201610694440.0A 2016-08-19 2016-08-19 A kind of low temperature resistant compost fermentation microbial inoculum with deodorization functions and application thereof Pending CN106244475A (en)

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CN107603920A (en) * 2017-11-02 2018-01-19 中山国晟生物工程有限公司 A kind of composite bacteria agent and its application in fermented organic waste increases numerous probiotics production biological organic fertilizer
CN107721469A (en) * 2017-10-27 2018-02-23 华南农业大学 The composite bacteria agent of ammonia emission and its application in a kind of reduction composting process
CN109679857A (en) * 2018-12-25 2019-04-26 广东高龙环保科技有限公司 A kind of compost maturity microbial inoculum
CN110423698A (en) * 2019-09-09 2019-11-08 山东庄氏农业科技有限公司 One plant of sporotrichum thermophile YM-2 and its composite bacteria agent and application
CN113354458A (en) * 2021-07-16 2021-09-07 珠海回田生物科技有限公司 Organic fertilizer and preparation method thereof
CN114933983A (en) * 2022-04-20 2022-08-23 湖南省微生物研究院 Microbial agent for synergistic emission reduction of livestock and poultry manure compost ammonia gas and greenhouse gas and preparation and application thereof
CN115160081A (en) * 2022-06-16 2022-10-11 西安智同天地环境工程有限公司 Composting method suitable for low-temperature environment
CN115611685A (en) * 2022-11-15 2023-01-17 山东泰山生力源集团股份有限公司 Microbial fermentation chicken manure compost as well as preparation method and application thereof
CN115611685B (en) * 2022-11-15 2024-04-30 山东泰山生力源集团股份有限公司 Microbial fermentation chicken manure compost and preparation method and application thereof

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CN105384480A (en) * 2015-11-16 2016-03-09 北京嘉博文生物科技有限公司 Biological humic acid organic material decomposition agent having deodorization function
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CN105733975A (en) * 2014-12-12 2016-07-06 河北省农林科学院遗传生理研究所 Multifunctional high efficient organic material decomposition agent and preparation method thereof
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Cited By (10)

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Publication number Priority date Publication date Assignee Title
CN107721469A (en) * 2017-10-27 2018-02-23 华南农业大学 The composite bacteria agent of ammonia emission and its application in a kind of reduction composting process
CN107603920A (en) * 2017-11-02 2018-01-19 中山国晟生物工程有限公司 A kind of composite bacteria agent and its application in fermented organic waste increases numerous probiotics production biological organic fertilizer
CN109679857A (en) * 2018-12-25 2019-04-26 广东高龙环保科技有限公司 A kind of compost maturity microbial inoculum
CN110423698A (en) * 2019-09-09 2019-11-08 山东庄氏农业科技有限公司 One plant of sporotrichum thermophile YM-2 and its composite bacteria agent and application
CN110423698B (en) * 2019-09-09 2020-09-22 山东庄氏农业科技有限公司 Thermophilic sporotrichum YM-2 and composite microbial agent and application thereof
CN113354458A (en) * 2021-07-16 2021-09-07 珠海回田生物科技有限公司 Organic fertilizer and preparation method thereof
CN114933983A (en) * 2022-04-20 2022-08-23 湖南省微生物研究院 Microbial agent for synergistic emission reduction of livestock and poultry manure compost ammonia gas and greenhouse gas and preparation and application thereof
CN115160081A (en) * 2022-06-16 2022-10-11 西安智同天地环境工程有限公司 Composting method suitable for low-temperature environment
CN115611685A (en) * 2022-11-15 2023-01-17 山东泰山生力源集团股份有限公司 Microbial fermentation chicken manure compost as well as preparation method and application thereof
CN115611685B (en) * 2022-11-15 2024-04-30 山东泰山生力源集团股份有限公司 Microbial fermentation chicken manure compost and preparation method and application thereof

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